Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo
- - - - -

DAB Peroxidase staining


  • Please log in to reply
3 replies to this topic

#1 baboon

baboon

    member

  • Active Members
  • Pip
  • 10 posts
0
Neutral

Posted 19 August 2009 - 11:14 AM

Hi,

I am trying to do IHC using an HRP-labeled secondary antibody. To detect the antibody I am using DAB + Nickel chloride as my substrate.
My question pertains to dissolving DAB substrate. I am trying to dissolve DAB in PBT (0.2% BSA, 0.1% triton X-100 in PBS) but it takes forever to dissolve it. Does anyone know a better way of how to dissolve DAB?

Thx.

#2 pesji

pesji

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 39 posts
0
Neutral

Posted 21 August 2009 - 03:45 AM

Hi,

I am trying to do IHC using an HRP-labeled secondary antibody. To detect the antibody I am using DAB + Nickel chloride as my substrate.
My question pertains to dissolving DAB substrate. I am trying to dissolve DAB in PBT (0.2% BSA, 0.1% triton X-100 in PBS) but it takes forever to dissolve it. Does anyone know a better way of how to dissolve DAB?

Thx.

Check the pH of your pBS it has to be 7.3, I would rather dissolve in pBS only

#3 baboon

baboon

    member

  • Active Members
  • Pip
  • 10 posts
0
Neutral

Posted 25 August 2009 - 09:46 AM

Thanks,
I shall try dissolving in PBS.....

#4 miraclestrain

miraclestrain

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 36 posts
0
Neutral

Posted 18 October 2009 - 08:13 PM

Hi,

I am trying to do IHC using an HRP-labeled secondary antibody. To detect the antibody I am using DAB + Nickel chloride as my substrate.
My question pertains to dissolving DAB substrate. I am trying to dissolve DAB in PBT (0.2% BSA, 0.1% triton X-100 in PBS) but it takes forever to dissolve it. Does anyone know a better way of how to dissolve DAB?

Thx.



Hi there,

I am using the same stuff, but i am using anti mouse HRP, for the DAB I am following the protocol provided by sigma, which is- Crush DAB- (atleast for 1 hour, to make it as a fine powder), try dissolving it in 3-5 ml warm ethanol, finally disolve it in PBS. I would like to discuss more about this, i am from Melbourne and you are?

MS




Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.